Andrographolide And Doxorubicin Liposomal Codelivery System For Treatment Of Breast Cancer Metastasis

Objective To explore the CPP-modified nano-drug delivery system for enhanced Doxorubicin delivery for anti-tumor activity and reduce the tumor migration and invasion,the Doxorubicin-Andrographolide loaded LMWP-modified were prepared.The delivery system provided a new method for the treatment of breast cancer metastasis.Methods The low molecular weight protamine(LMWP),a membrane reactive peptide,was modified onto the surface of liposomes by chemical synthesis.The liposome particle size and zeta potential were characterized by particle size analyzer,and analyzed its stability in vitro;by transmission electron microscopy(TEM)to observe the morphology and distribution of liposomes;the fluorescence imaging and flow cytometry were performed to determine cellular uptake efficiency;the proliferation inhibitory effect on the tumor cells was determined by the MTT method;wound-healing assay and cell migration and invasion assay were used to determine cell migration;the cell apoptosis activity was measured by using FACS to evaluate the efficacy of combination;and finally the mechanism of drug inhibition of tumor metastasis was investigated by Western blot assay.In this study,the tumor of mouse breast cancer cell line 4T1 xenograft mouse model as the research object,and the in vivo biological safety evaluation of liposome and drug delivery system,and the effects of the combined treatment system to inhibit tumor metastasis.Results The Lipo-CPP with diameter about 180 nm and zeta potential about 1 m V was obtained through formulation optimization.The TEM showed uniform sphericalbmorphology.The encapsulation efficiency of the Andro was 74.40 % and DOX 74.40 %.Liposomes remained stable within 72-hours,the particle size showed a very minor change in 37 °C in phosphate-buffered saline(PBS).The flow cytometric analysis showed that the Lipo-CPP increased intracellular delivery efficiency;The MTT assay showed that the Lipo-CPP with higher cytotoxicity to the tumor cells;wound healing and transwell migration assays indicated that the Lipo-CPP resulted in significantly decrease the ability of migration of 4T1 cells;the western blot indicated the VEGF was concerned with migration;in vivo results showed that the combination of DOX and Andro inhibited the metastasis efficiently.Conclusion The results of in vivo and in vitro experiments showed that enhanced anti-tumor activity both in vitro and in vivo.Modification of LMWP could improve the penetration ability of nanoparticles into the breast tumor cells and enhanced the intratumoral delivery.The CPP-modified nanoparticles could be a potential method for the treatment of breast resistant cancer.